Microarray analysis of 1,25(OH) 2D 3 regulated gene expression in human primary osteoblasts

Paola Tarroni, Isabella Villa, Emanuela Mrak, Francesca Zolezzi, Michela Mattioli, Claudio Gattuso, Alessandro Rubinacci

Research output: Contribution to journalArticlepeer-review

Abstract

Though extensive studies have been conducted, questions regarding the molecular effectors and pathways underlying the regulatory role of 1,25(OH) 2D 3 in human osteoblasts other than cell differentiation and matrix protein production remain unanswered. This study aims to identify genes and pathways that are modulated by 1,25(OH) 2D 3 treatment in human osteoblasts. Primary osteoblast cultures obtained from human bone tissue samples were treated with 1,25(OH) 2D 3 (10 -7 M) for 24 h and their transcritptomes were profiled by microarray analysis using the Affymetrix GeneChip®. Statistical analysis was conducted to identify genes whose expression is significantly modulated following 1,25(OH) 2D 3 treatment. One hundred and fifty-eight genes were found to be differentially expressed. Of these, 136 were upregulated, indicating clear transcriptional activation by 1,25(OH) 2D 3. Biostatistical evaluation of microarray data by Ingenuity Pathways Analysis (IPA) revealed a relevant modulation of genes involved in vitamin D metabolism (CYP24), immune functions (CD14), neurotransmitter transporters (SLC1A1, SLC22A3), and coagulation [thrombomodulin (THBD), tissue plasminogen activator (PLAT), endothelial protein C receptor (PROCR), thrombin receptor (F2R)]. We identified a restricted number of highly regulated genes and confirmed their differential expression by real-time quantitative PCR (RT qPCR). The present genome-wide microarray analysis on 1,25(OH) 2D 3-treated human osteoblasts reveals an interplay of critical regulatory and metabolic pathways and supports the hypothesis that 1,25(OH) 2D 3 can modulate the coagulation process through osteoblasts, activates osteoclastogenesis through inflammation signaling, modulates the effects of monoamines by affecting their reuptake.

Original languageEnglish
Pages (from-to)640-649
Number of pages10
JournalJournal of Cellular Biochemistry
Volume113
Issue number2
DOIs
Publication statusPublished - Feb 2012

Keywords

  • 1,25(OH) D
  • Human primary osteoblasts
  • Microarray
  • Transcriptional profiling

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

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